CN108096416A - 一种禽用抗支气管炎病毒的组合物、冻干粉及制备方法 - Google Patents

一种禽用抗支气管炎病毒的组合物、冻干粉及制备方法 Download PDF

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CN108096416A
CN108096416A CN201711292002.2A CN201711292002A CN108096416A CN 108096416 A CN108096416 A CN 108096416A CN 201711292002 A CN201711292002 A CN 201711292002A CN 108096416 A CN108096416 A CN 108096416A
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parts
extract
fowl
bronchitis virus
freeze
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杨兰明
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Chongqing Jinyuheng Technology Co Ltd
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Abstract

本发明公开了一种禽用抗支气管炎病毒的组合物，包括黄芪提取物1.2份，人参提取物1.5份、茯苓提取物1.7份，黄芪提取物1.8份，当归提取物1.5份，山药提取物2份，菊花提取物1.8份，淫羊霍提取物1.5份，大青叶提取物1.2份，黄连提取物1.2份，板蓝根提取物1.2份，麻黄提取物1.3份，白芍提取物1.1份，细辛提取物1.2份，五味子提取物1.3份。本发明采用黄芪提取物，人参提取物、茯苓提取物，黄芪提取物，当归提取物，白芍提取物提取物，细辛提取物提取物，五味子提取物是精选的采用对支气管炎病毒有阻断作用的药材，同时这些提取物也能够对受损后的细胞进行修复。能够有效治愈传染支气管炎病毒传播，增强鸡的免疫力。

Description

一种禽用抗支气管炎病毒的组合物、冻干粉及制备方法
技术领域
本发明涉及一种养殖技术领域，特别是涉及一种禽用抗支气管炎病毒的组合物、冻干粉及制备方法。
背景技术
病毒感染是一大类严重威胁禽畜健康的疾病，然而现在由于疫苗和抗病毒西药的广泛使用，导致很多类型的病毒发生了抗药性，禽类支气管炎是由支气管病毒引发的一种能够高度传染的呼吸道疾病，其为养殖过程带来较大的经济损失，实践中，通常采用提前接种疫苗或者西药完成预防病害的效果，但是这样容易使禽类产生抗药性，进而对消费者健康产生影响，现有技术中，缺少一种能够抗支气管炎病毒同时避免病毒产生抗药性的药物，使得能够安全地对抗支气管炎病毒进行防治。
因此本领域技术人员致力于开发一种能够抗病毒同时避免病毒产生抗药性的药物，使得能够安全地对抗支气管炎病毒进行防治。
发明内容
有鉴于现有技术的上述缺陷，本发明所要解决的技术问题是提供一种能够抗病毒同时避免病毒产生抗药性的药物，使得能够安全地对抗支气管炎病毒进行防治。
为实现上述目的，本发明提供了一种禽用抗支气管炎病毒的组合物，冻干粉、冻干粉的制备方法、及所述冻干粉作为饲料添加剂的应用，包括黄芪提取物1.2份，人参提取物1.5份、茯苓提取物1.7份，黄芪提取物1.8份，当归提取物1.5份，山药提取物2份，菊花提取物1.8份，淫羊霍提取物1.5份，大青叶提取物1.2份，黄连提取物1.2份，板蓝根提取物1.2份，麻黄提取物1.3份，白芍提取物1.1份，细辛提取物1.2份，五味子提取物1.3份。还包括禽用抗病毒卵黄抗体，禽用抗病毒转移因子1.8份和α-干扰素1.2份。所述禽用抗病毒卵黄抗体为抗鸡新城疫卵黄抗体，所述禽用抗病毒转移因子为抗鸡新城疫转移因子。
一种禽用抗支气管炎病毒的冻干粉，主要由以下重量份数的组分组成：黄芪提取物1.2份，人参提取物1.5份、茯苓提取物1.7份，黄芪提取物1.8份，当归提取物1.5份，山药提取物2份，菊花提取物1.8份，淫羊霍提取物1.5份，大青叶提取物1.2份，黄连提取物1.2份，板蓝根提取物1.2份，麻黄提取物1.3份，白芍提取物1.1份，细辛提取物1.2份，五味子提取物1.3份。
本发明通过选择这数10种中药材来进行禽用抗支气管炎的防治，是由于这10种中药材均具有提高禽体免疫能力的作用，同时能够有效抑制禽类支气管炎病毒。
这些药材均属于常见药材，可以通过在药材市场上购买取得。
较优的，还包括以下重量份数的组分：禽用抗病毒卵黄抗体70-80份，禽用抗病毒转移因子1.8份和α-干扰素1.2份。
较优的，还包括氨茶碱1.8份。
较优的，包括链霉素1份和以重量份计0.05％的多维生素。
一种禽用抗支气管炎病毒的冻干粉制备方法，包括以下步骤：
(1)分离毒株制备疫苗，灭活后制得乳油剂疫苗备用；
(2)中药提取液的制备：按照如权利要求1所述的重量配比，取相同份数的中药材，采用常规水煎制备所述中药材的提取液；
(3)将步骤(2)中的中药材提取液进行浓缩，浓缩至1g/ml的药液；
(4)将步骤(3)中的药液进行3000r/ml进行离心，离心20min，得上清液；
(5)将步骤(4)中的药液用0.22μm进行微孔滤膜过滤，高压蒸汽灭菌，4℃冷藏备用；
(6)免疫健康鸡群制备高免蛋：取步骤(1)制备的疫苗接种健康鸡群，当抗体效价达到1:256时，收集高免蛋清理、消毒后晾干备用；
(7)分离高免蛋卵黄：取步骤(6)制备的高免蛋分离卵黄，加入等体积的无菌水，充分搅拌至颜色发白的卵黄溶液，将卵黄溶液置于60-65℃的水浴中加热20min。再加入PH为4.9-5.2的预冷酸化水，加入量为卵黄溶液体积的6-8倍，混匀后室温下静置沉淀，取得上清液后离心分离；
(8)取步骤(7)离心后的上清液，过滤，使得滤液中禽用抗病毒卵黄抗体的干重为70-80份，在浓缩液中加入步骤(5)的灭菌药液，再加入氨茶碱1.8份，防腐剂0.01份，灭活剂0.2份，链霉素1份，耐热冻干保护剂3份，0.05％的维生素1份，混匀后预冻、冻干备用；
(9)以重量份数剂，在步骤(8)的干粉中加入禽用抗病毒转移因子1.8份和α-干扰素1.2份即可。
较优的，所述禽用抗支气管炎病毒转移因子的制备方法为：
(1)取刚屠宰的新鲜健康猪脾，用生理盐水反复冲洗后灭菌并去膜、脂肪、筋等；
(2)组织捣碎机以5000r/min的速度进行捣碎，共3次，每次间隔5min；
(3)冷却捣碎机，冷却后的细胞培养液以1500r/min的速度离心10min；
(4)去除离心液的上清液，续加氯化铵液，离心并去红细胞；
(5)利用小牛血清培养液制成猪脾细胞培养液；
(6)取步骤(5)中的猪脾细胞培养液100μl,加入质量浓度为100g/L的乙肝疫苗100μl；
(7)取步骤(6)中的猪脾细胞培养液,加入质量浓度为20g/L的支气管炎病毒疫苗50μl；
(8)将步骤(7)中的培养液加入蒸馏水1:1稀释，-25℃反复冻融6次，自来水融化后装入透析袋中；
(9)用1:4蒸馏水作为透析外液，4℃透析48h，取得透析液，4℃保存或冻干备用。
本发明中采用体外制备抗病因子的方法，比现有方法活体接疫苗后提取组织液的方法而言，其制备更具有经济性，且更容易制备，制备的抗病因子纯度也更高。
较优的，一种禽用抗支气管炎病毒冻干粉作为饲料添加剂的应用，包括以下步骤，将禽用抗支气管炎病毒冻干粉以1.2％的量加入禽用饲料中即可。
本发明的有益效果是：本发明采用黄芪提取物，人参提取物、茯苓提取物，黄芪提取物，当归提取物，山药提取物，菊花提取物，淫羊霍提取物，大青叶提取物，黄连提取物，板蓝根提取物，麻黄提取物，白芍提取物提取物，细辛提取物提取物，五味子提取物是精选的采用对支气管炎病毒有阻断作用的药材，同时这些提取物也能够对受损后的细胞进行修复。能够有效治愈传染支气管炎病毒传播，增强禽类的免疫力。
具体实施方式
下面结合实施例对本发明作进一步说明：
实施例1，一种禽用抗支气管炎病毒的组合物，包括黄芪提取物1.2份，人参提取物1.5份、茯苓提取物1.7份，黄芪提取物1.8份，当归提取物1.5份，山药提取物2份，菊花提取物1.8份，淫羊霍提取物1.5份，大青叶提取物1.2份，黄连提取物1.2份，板蓝根提取物1.2份，麻黄提取物1.3份，白芍提取物1.1份，细辛提取物1.2份，五味子提取物1.3份。
还包括禽用抗病毒卵黄抗体，禽用抗病毒转移因子1.8份和α-干扰素1.2份。禽用抗病毒卵黄抗体为抗鸡新城疫卵黄抗体，禽用抗病毒转移因子为抗鸡新城疫转移因子。
主要由以下重量份数的组分组成：黄芪提取物1.2份，人参提取物1.5份、茯苓提取物1.7份，黄芪提取物1.8份，当归提取物1.5份，山药提取物2份，菊花提取物1.8份，淫羊霍提取物1.5份，大青叶提取物1.2份，黄连提取物1.2份，板蓝根提取物1.2份，麻黄提取物1.3份，白芍提取物1.1份，细辛提取物1.2份，五味子提取物1.3份。
还包括以下重量份数的组分：禽用抗病毒卵黄抗体，禽用抗病毒转移因子80份和α-干扰素1.2份。还包括氨茶碱1.8份。
包括链霉素1份和以重量份计0.05％的多维生素。
本实施例中采用以下步骤来制备禽用抗支气管炎病毒的冻干粉制备方法，包括以下步骤：
(1)分离毒株制备疫苗，灭活后制得乳油剂疫苗备用；
(2)中药提取液的制备：按照如权利要求1的重量配比，取相同份数的中药材，采用常规水煎制备中药材的提取液；
(3)将步骤(2)中的中药材提取液进行浓缩，浓缩至1g/ml的药液；
(4)将步骤(3)中的药液进行3000r/ml进行离心，离心20min，得上清液；
(5)将步骤(4)中的药液用0.22μm进行微孔滤膜过滤，高压蒸汽灭菌，4℃冷藏备用；
(6)免疫健康鸡群制备高免蛋：取步骤(1)制备的疫苗接种健康鸡群，当抗体效价达到1:256时，收集高免蛋清理、消毒后晾干备用；
(7)分离高免蛋卵黄：取步骤(6)制备的高免蛋分离卵黄，加入等体积的无菌水，充分搅拌至颜色发白的卵黄溶液，将卵黄溶液置于60-65℃的水浴中加热20min。再加入PH为5.2的预冷酸化水，加入量为卵黄溶液体积的6.8倍，混匀后室温下静置沉淀，取得上清液后离心分离；(8)取步骤(7)离心后的上清液，过滤，使得滤液中禽用抗病毒卵黄抗体的干重为50-60份，在浓缩液中加入步骤(5)的灭菌药液，再加入氨茶碱1.8份，防腐剂0.01份，灭活剂0.2份，链霉素1份，耐热冻干保护剂3份，0.05％的维生素1份，混匀后预冻、冻干备用；(9)以重量份数剂，在步骤(8)的干粉中加入禽用抗病毒转移因子1.8份和α-干扰素1.2份即可。
一种禽用抗支气管炎病毒的冻干粉制备方法，禽用抗支气管炎病毒转移因子的制备方法为：
(1)取刚屠宰的新鲜健康猪脾，用生理盐水反复冲洗后灭菌并去膜、脂肪、筋等；
(2)组织捣碎机以5000r/min的速度进行捣碎，共3次，每次间隔5min；
(3)冷却捣碎机，冷却后的细胞培养液以1500r/min的速度离心10min；
(4)去除离心液的上清液，续加氯化铵液，离心并去红细胞；
(5)利用小牛血清培养液制成猪脾细胞培养液；
(6)取步骤(5)中的猪脾细胞培养液100μl,加入质量浓度为100g/L的乙肝疫苗100μl；
(7)取步骤(6)中的猪脾细胞培养液,加入质量浓度为20g/L的支气管炎病毒疫苗50μl；
(8)将步骤(7)中的培养液加入蒸馏水1:1稀释，-25℃反复冻融6次，自来水融化后装入透析袋中；
(9)用1:4蒸馏水作为透析外液，4℃透析48h，取得透析液，4℃保存或冻干备用。
一种禽用抗支气管炎病毒冻干粉作为饲料添加剂的应用，包括以下步骤，将禽用抗支气管炎病毒冻干粉以1.2％的量加入禽用饲料中即可。
试验方法：选取1月龄鸡200只，其中100只作为攻毒组隔离饲养，100只作为空白对照组隔离饲养，100作为试验组按照本发明的方式进行饲养，试验组采用实施例1制备的冻干粉，饲养方式为自由采食，自由饮水，24h后取试验组、对照组和攻毒组的鸡肌注接种稀释后的鸡支气管炎病毒液，感染后继续饲养，治疗时对照组和试验组每100ml注射液中分别含有冻干粉1g，采用肌注方式给药，0.1ml/kg体重，每日2次，攻毒组肌注生理盐水，用法同上，120h后得到表1结果如下：
组别 病鸡数/只 治愈数/只 有效数/只 治愈率/％
试验数 100 98 2 98％
攻毒组 100 18 10 /
空白对照组 0 / / /
由表1可知，本发明的治愈率为98％，治愈效果较好。同时保存容易，易被鸡群所接受。
以上详细描述了本发明的较佳具体实施例。应当理解，本领域的普通技术人员无需创造性劳动就可以根据本发明的构思作出诸多修改和变化。因此，凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的实验可以得到的技术方案，皆应在由权利要求书所确定的保护范围内。

Claims (10)

1.一种禽用抗支气管炎病毒的组合物，其特征是：包括黄芪提取物1.2份，人参提取物1.5份、茯苓提取物1.7份，黄芪提取物1.8份，当归提取物1.5份，山药提取物2份，菊花提取物1.8份，淫羊霍提取物1.5份，大青叶提取物1.2份，黄连提取物1.2份，板蓝根提取物1.2份，麻黄提取物1.3份，白芍提取物1.1份，细辛提取物1.2份，五味子提取物1.3份。
2.如权利要求1所述的禽用抗支气管炎病毒的组合物，其特征是：还包括禽用抗病毒卵黄抗体，禽用抗病毒转移因子1.8份和α-干扰素1.2份。
3.如权利要求1所述的禽用抗支气管炎病毒的组合物，其特征是：所述禽用抗病毒卵黄抗体为抗鸡新城疫卵黄抗体，所述禽用抗病毒转移因子为抗鸡新城疫转移因子。
4.一种禽用抗支气管炎病毒的冻干粉，其特征是：主要由以下重量份数的组分组成：黄芪提取物1.2份，人参提取物1.5份、茯苓提取物1.7份，黄芪提取物1.8份，当归提取物1.5份，山药提取物2份，菊花提取物1.8份，淫羊霍提取物1.5份，大青叶提取物1.2份，黄连提取物1.2份，板蓝根提取物1.2份，麻黄提取物1.3份，白芍提取物1.1份，细辛提取物1.2份，五味子提取物1.3份。
5.根据权利要求4所述禽用抗支气管炎病毒的冻干粉，其特征是：还包括以下重量份数的组分：禽用抗病毒卵黄抗体70-80份，禽用抗病毒转移因子1.8份和α-干扰素1.2份。
6.根据权利要求4或5所述的禽用抗支气管炎病毒的冻干粉，其特征是：还包括氨茶碱1.8份。
7.如权要求4或5所述的禽用抗支气管炎病毒的冻干粉，其特征是：包括链霉素1份和以重量份计0.05％的多维生素。
8.一种禽用抗支气管炎病毒的冻干粉制备方法，其特征在于：包括以下步骤：
(1)分离毒株制备疫苗，灭活后制得乳油剂疫苗备用；
(2)中药提取液的制备：按照如权利要求1所述的重量配比，取相同份数的中药材，采用常规水煎制备所述中药材的提取液；
(3)将步骤(2)中的中药材提取液进行浓缩，浓缩至1g/ml的药液；
(4)将步骤(3)中的药液进行3000r/ml进行离心，离心20min，得上清液；
(5)将步骤(4)中的药液用0.22μm进行微孔滤膜过滤，高压蒸汽灭菌，4℃冷藏备用；
(6)免疫健康鸡群制备高免蛋：取步骤(1)制备的疫苗接种健康鸡群，当抗体效价达到1:256时，收集高免蛋清理、消毒后晾干备用；
(7)分离高免蛋卵黄：取步骤(6)制备的高免蛋分离卵黄，加入等体积的无菌水，充分搅拌至颜色发白的卵黄溶液，将卵黄溶液置于60-65℃的水浴中加热20min。再加入PH为4.9-5.2的预冷酸化水，加入量为卵黄溶液体积的6-8倍，混匀后室温下静置沉淀，取得上清液后离心分离；
(8)取步骤(7)离心后的上清液，过滤，使得滤液中禽用抗病毒卵黄抗体的干重为70-80份，在浓缩液中加入步骤(5)的灭菌药液，再加入氨茶碱1.8份，防腐剂0.01份，灭活剂0.2份，链霉素1份，耐热冻干保护剂3份，0.05％的维生素1份，混匀后预冻、冻干备用；
(9)以重量份数剂，在步骤(8)的干粉中加入禽用抗病毒转移因子1.8份和α-干扰素1.2份即可。
9.如权利要求8所述的一种禽用抗支气管炎病毒的冻干粉制备方法，其特征是：所述禽用抗支气管炎病毒转移因子的制备方法为：
(1)取刚屠宰的新鲜健康猪脾，用生理盐水反复冲洗后灭菌并去膜、脂肪、筋等；
(2)组织捣碎机以5000r/min的速度进行捣碎，共3次，每次间隔5min；
(3)冷却捣碎机，冷却后的细胞培养液以1500r/min的速度离心10min；
(4)去除离心液的上清液，续加氯化铵液，离心并去红细胞；
(5)利用小牛血清培养液制成猪脾细胞培养液；
(6)取步骤(5)中的猪脾细胞培养液100μl,加入质量浓度为100g/L的乙肝疫苗100μl；
(7)取步骤(6)中的猪脾细胞培养液,加入质量浓度为20g/L的支气管炎病毒疫苗50μl；
(8)将步骤(7)中的培养液加入蒸馏水1:1稀释，-25℃反复冻融6次，自来水融化后装入透析袋中；
(9)用1:4蒸馏水作为透析外液，4℃透析48h，取得透析液，4℃保存或冻干备用。
10.一种如权利要求4或5所述的一种禽用抗支气管炎病毒冻干粉作为饲料添加剂的应用，其特征是:包括以下步骤，将禽用抗支气管炎病毒冻干粉以1.2％的量加入禽用饲料中即可。
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